线粒体DNA突变与大鼠乙醇性肝纤维化的关系

The relationship between mitochondria DNA mutation and alcoholic liver damage in rats

目的对线粒体DNA ATP酶6,8基因(mtDNA ATPase 6,8gene)突变与乙醇性肝损伤发生发展之间的关系进行研究,从一个新的角度对乙醇性肝病(ALD)的发病机制进行探讨。方法应用乙醇加橄榄油灌胃的方式建立大鼠乙醇性肝损伤动物模型并设对照组(等量生理盐水),成模后进行肝脏组织学观察,PCR扩增各组线粒体DNA ATPase 6,8基因并进行序列测定。以生物发光法分别测定各组肝组织中ATP含量,判定ATPase 6,8基因突变对肝细胞内ATP能量合成的影响。结果模型组大鼠肝细胞线粒体出现形态学的改变;线粒体DNA ATPase 6亚基基因发现两处点突变;对照组未检出突变。模型组大鼠肝细胞内ATP含量明显降低(P<0.05)。结论持续过量的乙醇摄入可造成肝细胞线粒体形态和mtDNA ATPase基因突变,这可能是乙醇性肝损伤的重要发病机制。

Objective To investigate the relationship between the development of alcoholic liver damage and the mtDNA ATPase 6,8 gene mutation. Methods By use the method of alcohol infusion to the stomach of rats and olive oil balanced diet to make animal models of alcoholic liver damage, and the same route for saline control group. The blood samples and liver tissue of each group was collected. The mtDNA ATPase 6,8 genes were amplified by PCR and then the nucleotide sequence of the amplified products were detected. The ATP content of the liver tissue of each group was detected by bioluminescence method. Results The morphological changes of mitochondria were found and two point mutations were detected at mtDNA ATPase 6 gene in the alcoholic liver damage group, they were T8151C and C8300T, and No mutation site was detected in the control group.The ATP content of liver ceils in alcoholic liver damage group was markedly lower than the control group. Conclusion Alcohol may damage mitochondria and cause mutation of the mtDNA ATPase 6,8 genes, This may be one of the important pathogenesis of the alcoholic liver disease.